title: Characterization of squamous cell carcinoma in an organotypic culture via subsurface non-linear optical molecular imaging.
abstract: Tristetraprolin (TTP) is an RNA-binding protein which downregulates multiple cytokines that mediate progression of head and neck squamous cell carcinoma (HNSCC). We previously showed that HNSCC cells with shRNA-mediated knockdown of TTP are more invasive than controls. In this study, we use control and TTP-deficient cells to present a novel subsurface non-linear optical molecular imaging method using a three-dimensional (3D) organotypic construct, and compare the live cell imaging data to histology of fixed tissue specimens. This manuscript describes how to prepare and image the novel organotypic system that closely mimics HNSCC in a clinical setting. The oral cancer equivalent (OCE) system allows HNSCC cells to stratify and invade beyond the basement membrane into underlying connective tissue prepared from decellularized human dermal tissue. The OCE model was inspired by tissue engineering strategies to prepare autologous transplants from human keratinocytes. Advantages of this method over previously used in vitro cancer models include the simulation of the basement membrane and complex connective tissue in the construct, in addition to the ability to track the 3D movement of live invading cells and quantify matrix destruction over time. The OCE model and novel live cell imaging strategy may be applied to study other types of 3D tissue constructs.
authors.affiliation: Department of Periodontics and Oral Medicine, School of Dentistry, University of Michigan, MI 48109-1078, USA.
authors.forename: Christina S
authors.initials: CS
authors.lastname: Scanlon
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Van Tubergen
authors.forename: Leng-Chun
authors.initials: LC
authors.lastname: Chen
authors.forename: Sakib F
authors.initials: SF
authors.lastname: Elahi
authors.forename: Shiuhyang
authors.initials: S
authors.lastname: Kuo
authors.forename: Stephen
authors.initials: S
authors.lastname: Feinberg
authors.forename: Mary-Ann
authors.initials: MA
authors.lastname: Mycek
authors.forename: Nisha J
authors.initials: NJ
authors.lastname: D'Silva
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D018929:Cell Culture Techniques; D015972:Gene Expression Regulation, Neoplastic; D006258:Head and Neck Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000473:pathology; D061848:Optical Imaging / Q000379:methods; D023822:Tissue Engineering / Q000379:methods; D051816:Tristetraprolin / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D051816:Tristetraprolin
keywords: Head and neck cancer; bioimaging; invasion; tissue engineering; translational research